[12-Aug-2008] PRIMER DESIGNINGStep by Step Approach to Designing of Primers using NCBI PRIMER BLAST Approach

Neha MittalFOREST RESEARCH INSTITUTEDEHRADUN

Primer Designing Using Primer BLAST:

Go to the NCBI (National Centre for Biotechnology Information) site at www.ncbi.nlm.nih.gov

Search the all databases for particular species whose primers has to be design. Put the name of species in upper column and then click on search option.

Click on to the nucleotide sequences: Nucleotide (DNA and RNA sequences), Genome Survey Sequences and EST sequences, from which primer(s) has to be design.

Click on the particular link of sequenced genome

Click on the link and Open the seuqence

Click on the option from Right Hand Side Bar: Analyze this Sequence for Pick Primers

PRIMER-BLAST will open

Paste either accession no. or copy, paste the FASTA format sequence of DNA in the PCR Template Blank Space or upload the FASTA file in Browse Option.

Completely fill the Primer Parameters as per kind of primer requirements.

Click on the option Get Primers

The complete avilable sequence of primers that matches for provided sequence will get display in the following order/format